Molecular cloning, characterisation and expression of two catalase genes from peach

2004 ◽  
Vol 31 (4) ◽  
pp. 349 ◽  
Author(s):  
Francesca Bagnoli ◽  
Susanna Danti ◽  
Valentina Magherini ◽  
Radiana Cozza ◽  
Anna M. Innocenti ◽  
...  
Keyword(s):  
Stress Responses ◽  
Seasonal Changes ◽  
Prunus Persica ◽  
Vascular Tissue ◽  
In Situ Hybridisation ◽  
Leaf Tissue ◽  
Amino Acid Sequences ◽  
Cdna Clones

Two cDNA clones encoding catalase (Cat1 and Cat2) from peach [Prunus persica (L.) Batsch] were identified, that show homologies to other plant catalases. The nucleotide sequences of the two coding regions showed 88% identity to each other. The amino acid sequences predicted from the two full-length clones showed the highest homology to a catalase from cotton and Nicotiana plumbaginifolia L. and included C-terminal tri-peptides typical of those used to target proteins to peroxisomes. Southern hybridisation analysis suggested the existence of two catalase genes in peach. The expression of Cat1 and Cat2 was determined in seeds, vegetative tissue, leaves during the seasonal cycle and in leaves in response to light / dark treatments. Cat1 had high levels of expression only in leaf tissue and was responsive to light and seasonal changes. Cat2 had high levels of expression in in vitro shoots and was also responsive to seasonal changes, but not to light. In situ hybridisations to leaf tissue indicated that the expression of Cat1 was localised mainly in palisade cells, while Cat2 mRNA was present in the vascular tissue. The results of the expression analysis and in situ hybridisation suggest a role for Cat1 in photorespiration and for Cat2 in stress responses.

scholarly journals Synaptopodin: An Actin-associated Protein in Telencephalic Dendrites and Renal Podocytes

1997 ◽  
Vol 139 (1) ◽  
pp. 193-204 ◽  
Author(s):  
Peter Mundel ◽  
Hans W. Heid ◽  
Thomas M. Mundel ◽  
Meike Krüger ◽  
Jochen Reiser ◽  
...  
Keyword(s):  
Dendritic Spines ◽  
Cultured Cells ◽  
Rat Kidney ◽  
Amino Acid Sequences ◽  
Cdna Clones ◽  
Globular Domain ◽  
Postnatal Maturation ◽  
Human And Mouse

Synaptopodin is an actin-associated protein of differentiated podocytes that also occurs as part of the actin cytoskeleton of postsynaptic densities (PSD) and associated dendritic spines in a subpopulation of exclusively telencephalic synapses. Amino acid sequences determined in purified rat kidney and forebrain synaptopodin and derived from human and mouse brain cDNA clones show no significant homology to any known protein. In particular, synaptopodin does not contain functional domains found in receptor-clustering PSD proteins. The open reading frame of synaptopodin encodes a polypeptide with a calculated Mr of 73.7 kD (human)/74.0 kD (mouse) and an isoelectric point of 9.38 (human)/9.27 (mouse). Synaptopodin contains a high amount of proline (∼20%) equally distributed along the protein, thus virtually excluding the formation of any globular domain. Sequence comparison between human and mouse synaptopodin revealed 84% identity at the protein level. In both brain and kidney, in vivo and in vitro, synaptopodin gene expression is differentiation dependent. During postnatal maturation of rat brain, synaptopodin is first detected by Western blot analysis at day 15 and reaches maximum expression in the adult animal. The exclusive synaptopodin synthesis in the telencephalon has been confirmed by in situ hybridization, where synaptopodin mRNA is only found in perikarya of the olfactory bulb, cerebral cortex, striatum, and hippocampus, i.e., the expression is restricted to areas of high synaptic plasticity. From these results and experiments with cultured cells we conclude that synaptopodin represents a novel kind of proline-rich, actin-associated protein that may play a role in modulating actin-based shape and motility of dendritic spines and podocyte foot processes.

Structure and expression of ferritin genes in a human promyelocytic cell line that differentiates in vitro

1986 ◽  
Vol 6 (2) ◽  
pp. 566-573
Author(s):  
C C Chou ◽  
R A Gatti ◽  
M L Fuller ◽  
P Concannon ◽  
A Wong ◽  
...  
Keyword(s):  
Cell Line ◽  
Terminal Differentiation ◽  
Amino Acid Sequences ◽  
Multigene Families ◽  
Cdna Clones ◽  
Macrophage Differentiation ◽  
Time Points ◽  
Human Ferritin ◽  
Complex Manner

HL-60 is a human promyelocytic cell line with the capability of differentiating in vitro to give neutrophils, macrophages, or eosinophils. We screened libraries of HL-60 cDNA clones representing different time points during these differentiation processes to isolate clones corresponding to mRNAs whose expression is regulated during terminal differentiation. Upon sequencing this group of regulated clones, one clone encoding the heavy subunit and two clones encoding the light subunit of human ferritin were identified by reference to published amino acid sequences. Southern blot analyses showed that these clones are encoded by distinct multigene families. These clones identify two mRNAs whose ratios vary in a complex manner during both neutrophil and macrophage differentiation.

scholarly journals Potential and restrictions of Poincianella pyramidalis (Tul.) L. P. Queiroz as native forage in the Brazilian semi-arid region

2019 ◽  
Vol 42 ◽  
pp. e47460
Author(s):  
Andrezza Araújo de França ◽  
Divan Soares da Silva ◽  
Josean Tavares Fechine ◽  
Francinilda Alves de Sousa ◽  
Alberício Pereira de Andrade ◽  
...  
Keyword(s):  
Leaf Age ◽  
Plant Defence ◽  
Leaf Tissue ◽  
Secondary Compounds ◽  
Mineral Matter ◽  
Endemic Plant ◽  
Leaf Maturity ◽  
13C Nuclear Magnetic Resonance

Poincianella pyramidalis (catingueira) is a endemic plant of the Caatinga, selected by animals grazing on native pasture. With the aim of evaluating characteristics indicative of its nutritional quality, 10 plants were selected and identified, sampled at five different ages, were used to determine dry matter (DM), crude protein (CP), neutral detergent fibre (NDF), mineral matter (MM), DM degradability (Deg DM), NDF degradability (Deg NDF) and in situ and in vitro leaf-tissue degradability. Phytochemical prospection was performed, and 1H and 13C nuclear magnetic resonance applied to detect the presence of secondary compounds. The data were submitted to analysis of variance and Tukey’s test at 5%, and correlation analysis was carried out on the variables for leaf maturity in days. The levels of CP, NDF and Deg NDF showed a negative correlation with the increases in leaf age. Leaf-tissue degradation was restricted due to a physical barrier developed in the leaf fragments, which can be attributed to plant defence mechanisms. The in situ degradability of the cell wall components decreased with the increase in leaf age. The high levels of tannins and lignin, and the strong presence of flavonoids, should be considered for their anti-nutritional and pharmacological potential.

scholarly journals Molecular characterization of Penicillium chrysogenum virus: reconsideration of the taxonomy of the genus Chrysovirus

2004 ◽  
Vol 85 (7) ◽  
pp. 2111-2121 ◽  
Author(s):  
Daohong Jiang ◽  
Said A. Ghabrial
Keyword(s):  
Molecular Characterization ◽  
Penicillium Chrysogenum ◽  
Sequence Similarity ◽  
Minor Component ◽  
Amino Acid Sequences ◽  
Nucleotide Sequencing ◽  
Dsrna Segment ◽  
Cdna Clones

Molecular cloning and complete nucleotide sequencing of Penicillium chrysogenum virus (PcV) dsRNAs indicated that PcV virions contained four dsRNA segments with sizes of 3562, 3200, 2976 and 2902 bp. Each dsRNA segment had unique sequences and contained a single large open reading frame (ORF). In vitro translation of transcripts derived from full-length cDNA clones of PcV dsRNAs yielded single products of sizes similar to those predicted from the deduced amino acid sequences of the individual ORFs. Sequence similarity searches revealed that dsRNA1 encodes a putative RNA-dependent RNA polymerase. In this study, it was determined that dsRNA2 encodes the major capsid protein and that p4, encoded by dsRNA4, is virion-associated as a minor component. All four dsRNAs of PcV, like the genomic segments of viruses with multipartite genomes, were found to have extended regions of highly conserved terminal sequences at both ends. In addition to the strictly conserved 5′-terminal 10 nt, a second region consisting of reiteration of the sequence CAA was found immediately upstream of the AUG initiator codon. These (CAA) n repeats are reminiscent of the translational enhancer elements of tobamoviruses. The 3′-terminal 14 nt were also strictly conserved. As PcV and related viruses with four dsRNA segments (genus Chrysovirus) have not been previously characterized at the molecular level, they were provisionally classified in the family Partitiviridae, comprising viruses with bipartite genomes. This study represents the first report on molecular characterization of a chrysovirus and the results suggest the creation of a new family of mycoviruses with multipartite dsRNA genomes to accommodate PcV and related viruses.

scholarly journals ADAPTIVE CAPACITY OF SOME LAVENDER AND LAVANDIN CULTIVARSIN VITRO AND IN SITU

AGROFOR ◽  
2018 ◽  
Vol 2 (1) ◽  
Author(s):  
Oksana GREBENNIKOVA ◽  
Anfisa PALIY ◽  
Valentina BRAILKO ◽  
Olga MITROFANOVA ◽  
Valery RABOTYAGOV ◽  
...  
Keyword(s):  
Phenolic Compounds ◽  
Photosynthetic Activity ◽  
Bound Water ◽  
Leaf Tissue ◽  
Viral Pathogens ◽  
Intact Plants ◽  
Seed Propagation ◽  
Proline Concentration

Lavandula angustifolia Mill. and (LavandulaxintermediaEmericexLoisel) arepromising fragrant plants with medicinal, aromatic and ornamental properties.Since the collection plantations of these crops are very damaged with viralpathogens and there is lack of seed propagation in valuable cultivars 'Belyanka','Record' (lavender) and 'Rabat', 'Snezhnyi Bars' (lavandin), were introduced invitro. Chemotherapy was used for cleaning up. Regenerants were cultured (4-5months) on MS medium with 0. 3 mg L- Kinetin, 0. 025 mg L- NAA and 0. 25 mgL- GA3 at 25±1°C under 16-h photoperiod. Intact plants were studied during thegrowing season. In order to reveal plants` biotechnological and genetic capacitysome biochemical stress indicators, indexes of photosynthetic activity and waterregime were identified. Under the open field cultivation, tested plants were rich inascorbic acid, phenolic compounds, and redox enzymes (catalase, polyphenoloxidase, superoxide dismutase) were active. Leaf tissue hydration was 56-62%,with greater part of bound water. Photosynthetic activity was reduced only in thesamples with visible damages with viral pathogens. In plants cultured in vitro,amount of ascorbic acid and phenolic compounds were lower, so as enzymaticactivity and proline concentration were higher than in intact plants. The rate ofhydration was high (70-77%), with the same trend of water fractional composition.Photosynthetic activity and vitality index indicated no photoinhibition. It wasfound out the lavandin cultivars had better capacity for a wide use under variousculture conditions.

Human acidic ribosomal phosphoproteins P0, P1, and P2: analysis of cDNA clones, in vitro synthesis, and assembly

1987 ◽  
Vol 7 (11) ◽  
pp. 4065-4074
Author(s):  
B E Rich ◽  
J A Steitz
Keyword(s):  
Amino Acid ◽  
Cross Reactivity ◽  
Amino Acid Sequences ◽  
T7 Rna Polymerase ◽  
Cdna Clones ◽  
In Vitro Synthesis ◽  
E Coli ◽  
P Proteins ◽  
Carboxy Terminal

cDNA clones encoding three antigenically related human ribosomal phosphoproteins (P-proteins) P0, P1, and P2 were isolated and sequenced. P1 and P2 are analogous to Escherichia coli ribosomal protein L7/L12, and P0 is likely to be an analog of L10. The three proteins have a nearly identical carboxy-terminal 17-amino-acid sequence (KEESEESD(D/E)DMGFGLFD-COOH) that is the basis of their immunological cross-reactivity. The identities of the P1 and P2 cDNAs were confirmed by the strong similarities of their encoded amino acid sequences to published primary structures of the homologous rat, brine shrimp, and Saccharomyces cerevisiae proteins. The P0 cDNA was initially identified by translation of hybrid-selected mRNA and immunoprecipitation of the products. To demonstrate that the coding sequences are full length, the P0, P1, and P2 cDNAs were transcribed in vitro by bacteriophage T7 RNA polymerase and the resulting mRNAs were translated in vitro. The synthetic P0, P1, and P2 proteins were serologically and electrophoretically identical to P-proteins extracted from HeLa cells. These synthetic P-proteins were incorporated into 60S but not 40S ribosomes and also assembled into a complex similar to that described for E. coli L7/L12 and L10.

Nucleotide (cDNA) sequence encoding the horse gonadotrophin α-subunit

1987 ◽  
Vol 115 (2) ◽  
pp. 341-346 ◽  
Author(s):  
F. Stewart ◽  
J. A. Thomson ◽  
S. E. A. Leigh ◽  
J. M. Warwick
Keyword(s):  
Amino Acid ◽  
Peptide Sequencing ◽  
Amino Acid Sequences ◽  
In Vitro Translation ◽  
Northern Hybridization ◽  
Cdna Clones ◽  
Subunit Gene ◽  
Α Subunit ◽  
Pituitary Glands

ABSTRACT Several cDNA clones corresponding to mRNA for the α-subunit of the horse (Equus caballus) pituitary and placental (chorionic) gonadotrophic hormones have been isolated and sequenced. Polyadenylated mRNA was purified from horse pituitary glands (the source of FSH and LH) and horse placental tissues (the source of chorionic gonadotrophin; CG). The mRNA preparations were characterized by in-vitro translation and Northern hybridization techniques using human and ovine gonadotrophin cDNA clones as probes. Complementary DNA libraries were created from the pituitary and placental mRNAs and a human CG α-subunit probe was used to isolate several horse α-subunit cDNA clones. The α-subunit nucleotide sequence from both sources of tissue was identical, thereby indicating that in the horse (as in man) the same gonadotrophin α-subunit gene is expressed in the pituitary and placenta. Our results are consistent with transcription of a single α-subunit gene for all the glycoprotein hormones in the horse, and we suggest that the reported differences between the horse CG and FSH α-subunit amino acid sequences determined by conventional peptide sequencing methods arose due to errors in the FSH α-subunit sequence. Comparison of the deduced amino acid sequence of the horse α-subunit with that of other α-subunit sequences indicated a number of significant differences which may be related to the unusual receptor-binding properties of the equine gonadotrophins. J. Endocr. (1987) 115, 341–346

Immunological mechanisms to establish embryo tolerance in early bovine pregnancy

2011 ◽  
Vol 23 (5) ◽  
pp. 619 ◽  
Author(s):  
A. E. Groebner ◽  
K. Schulke ◽  
J. C. Schefold ◽  
G. Fusch ◽  
F. Sinowatz ◽  
...  
Keyword(s):  
In Situ Hybridisation ◽  
Immunohistochemical Analysis ◽  
Kynurenine Pathway ◽  
Endometrial Cells ◽  
Maternal Rejection ◽  
Immunological Mechanisms ◽  
Bovine Pregnancy ◽  
Coculture Model

A well-balanced immunological interaction between mother and the semi-allogenic embryo is of particular importance. The objective of the present study was to analyse mechanisms of immune tolerance in bovine pregnancy during peri-implantation. Simmental heifers inseminated with either cryopreserved spermatozoa or seminal plasma were killed 12, 15 or 18 days after oestrus. Uteri were flushed for the recovery of conceptuses and the ipsilateral intercaruncular endometrium was sampled for gene expression analysis. Indoleamine 2,3-dioxygenase (IDO) mRNA, coding for the initial enzyme of the kynurenine pathway, was 18-fold (P < 0.001) more abundant in the endometrium of Day 18 pregnant v. non-pregnant animals. Tandem mass spectrometry revealed a decrease of endometrial l-tryptophan (P = 0.0008), but an increase of l-kynurenine concentration (P = 0.005) from Day 12 to Day 18, suggesting increasing IDO activity (P < 0.03). An in vitro coculture model of endometrial cells showed an induction of IDO expression following interferon-τ exposure primarily in stroma cells, which was confirmed by in situ hybridisation localising IDO mRNA mainly in deep stroma cells. Immunohistochemical analysis revealed fewer CD45-positive leucocytes in the zona basalis of pregnant animals. Elevated IDO activity may reduce the presence of leucocytes in the pregnant endometrium, providing a possible mechanism for protecting the semi-allogenic conceptus from maternal rejection.

scholarly journals Human acidic ribosomal phosphoproteins P0, P1, and P2: analysis of cDNA clones, in vitro synthesis, and assembly.

1987 ◽  
Vol 7 (11) ◽  
pp. 4065-4074 ◽  
Author(s):  
B E Rich ◽  
J A Steitz
Keyword(s):  
Amino Acid ◽  
Cross Reactivity ◽  
Amino Acid Sequences ◽  
T7 Rna Polymerase ◽  
Cdna Clones ◽  
In Vitro Synthesis ◽  
E Coli ◽  
P Proteins ◽  
Carboxy Terminal

cDNA clones encoding three antigenically related human ribosomal phosphoproteins (P-proteins) P0, P1, and P2 were isolated and sequenced. P1 and P2 are analogous to Escherichia coli ribosomal protein L7/L12, and P0 is likely to be an analog of L10. The three proteins have a nearly identical carboxy-terminal 17-amino-acid sequence (KEESEESD(D/E)DMGFGLFD-COOH) that is the basis of their immunological cross-reactivity. The identities of the P1 and P2 cDNAs were confirmed by the strong similarities of their encoded amino acid sequences to published primary structures of the homologous rat, brine shrimp, and Saccharomyces cerevisiae proteins. The P0 cDNA was initially identified by translation of hybrid-selected mRNA and immunoprecipitation of the products. To demonstrate that the coding sequences are full length, the P0, P1, and P2 cDNAs were transcribed in vitro by bacteriophage T7 RNA polymerase and the resulting mRNAs were translated in vitro. The synthetic P0, P1, and P2 proteins were serologically and electrophoretically identical to P-proteins extracted from HeLa cells. These synthetic P-proteins were incorporated into 60S but not 40S ribosomes and also assembled into a complex similar to that described for E. coli L7/L12 and L10.

scholarly journals Novel lncRNA UPLA1 mediates tumorigenesis and prognosis in lung adenocarcinoma

2020 ◽  
Vol 11 (11) ◽  
Author(s):  
Xiaoyang Han ◽  
Hua Jiang ◽  
Jianni Qi ◽  
Jiamei Li ◽  
Jinghan Yang ◽  
...  
Keyword(s):  
Lung Adenocarcinoma ◽  
In Situ Hybridisation ◽  
Vital Role ◽  
Luciferase Reporter ◽  
Fluorescence In Situ Hybridisation ◽  
Biological Functions ◽  
Pulldown Assay

AbstractWith the development of molecular biotechnology and sequencing techniques, long non-coding RNAs (lncRNAs) have been shown to play a vital role in a variety of cancers including lung cancer. In our previous study, we used RNA sequencing and high-content screening proliferation screening data to identify lncRNAs that were significantly associated with tumour biological functions such as LINC01426. Herein, based on previous work, we report a novel lncRNA UPLA1 (upregulation promoting LUAD-associated transcript-1), which has not been explored or reported in any previous studies. Our results showed that UPLA1 is highly expressed and regulates important biological functions in lung adenocarcinoma. In vitro experiments revealed that UPLA1 promoted the migration, invasion, and proliferation abilities, and is related to cell cycle arrest, in lung adenocarcinoma cells. Moreover, the upregulation of UPLA1 significantly improved the growth of tumours in vivo. We identified that UPLA1 was mainly located in the nucleus using fluorescence in situ hybridisation, and that it promoted Wnt/β-catenin signalling by binding to desmoplakin using RNA pulldown assay and mass spectrometry. Additionally, luciferase reporter assay revealed that YY1 is the transcription factor of UPLA1 and suppressed the expression of UPLA1 as a transcriptional inhibitor. This finding provides important evidence regarding the two roles of YY1 in cancer. Furthermore, in situ hybridisation assay results showed that UPLA1 was closely related to the prognosis and tumour, node, metastasis (TNM) stage of lung adenocarcinoma. In summary, our results suggest that the novel lncRNA UPLA1 promotes the progression of lung adenocarcinoma and may be used as a prognostic marker, and thus, has considerable clinical significance.

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